The impact of physiological noise on hemodynamic-derived estimates of directed functional connectivity

This work was supported by a grant of the BrainLinks-BrainTools Cluster of Excellence funded by the German Research Foundation (DFG, Grant Number EXC 1086).Peer reviewedPostprintPostprin

PHIP-associated Chung-Jansen syndrome: Report of 23 new individuals

In 2016 and 2018, Chung, Jansen and others described a new syndrome caused by haploinsufficiency of PHIP (pleckstrin homology domain interacting protein, OMIM *612,870) and mainly characterized by developmental delay (DD), learning difficulties/intellectual disability (ID), behavioral abnormalities, facial dysmorphism and obesity (CHUJANS, OMIM #617991). So far, PHIP alterations appear to be a rare cause of DD/ID. “Omics” technologies such as exome sequencing or array analyses have led to the identification of distinct types of alterations of PHIP, including, truncating variants, missense substitutions, splice variants and large deletions encompassing portions of the gene or the entire gene as well as adjacent genomic regions. We collected clinical and genetic data of 23 individuals with PHIP-associated Chung-Jansen syndrome (CHUJANS) from all over Europe. Follow-up investigations (e.g. Sanger sequencing, qPCR or Fluorescence-in-situ-Hybridization) and segregation analysis showed either de novo occurrence or inheritance from an also (mildly) affected parent. In accordance with previously described patients, almost all individuals reported here show developmental delay (22/23), learning disability or ID (22/23), behavioral abnormalities (20/23), weight problems (13/23) and characteristic craniofacial features (i.e. large ears/earlobes, prominent eyebrows, anteverted nares and long philtrum (23/23)). To further investigate the facial gestalt of individuals with CHUJANS, we performed facial analysis using the GestaltMatcher approach. By this, we could establish that PHIP patients are indistinguishable based on the type of PHIP alteration (e.g. missense, loss-of-function, splice site) but show a significant difference to the average face of healthy individuals as well as to individuals with Prader-Willi syndrome (PWS, OMIM #176270) or with a CUL4B-alteration (Intellectual developmental disorder, X-linked, syndromic, Cabezas type, OMIM #300354). Our findings expand the mutational and clinical spectrum of CHUJANS. We discuss the molecular and clinical features in comparison to the published individuals. The fact that some variants were inherited from a mildly affected parent further illustrates the variability of the associated phenotype and outlines the importance of a thorough clinical evaluation combined with genetic analyses for accurate diagnosis and counselling

In Vitro Anti-inflammatory Effects of Equisetum Arvense Are Not Solely Mediated by Silica

Equisetum arvense, known as common horsetail, is used for the treatment of inflammatory diseases and is the plant with the highest concentration of silica. Yet it is unknown if the medicinal properties are mediated by its silica content. In the current study, optimal conditions for silica-rich horsetail preparations were identified. Bioactivity of the preparations was analyzed in vitro using flow cytometry-based activity and functionality profiling of primary human lymphocytes as well as cytokine measurement using a classical ELISA technique. Experiments revealed that horsetail preparations suppress activation and proliferation of lymphocytes by an interleukin-2-dependent mechanism. The effect increased with the silica concentration in the decoctions. Lymphocytesʼ polyfunctionality was also influenced, shown by a downregulation of IFN-γ. Analytical profiling by HPLC-UV-MS and bioactivity testing revealed relevant immunosuppressive concentrations of a component that has been identified as isoquercitrin. Our results show that both silica and isoquercitrin are active compounds of horsetail preparations

Investigations on the constitutional types under consideration of anthropometric data, autonomic regulation and immunological parameters

Over time different systems were developed for the characterization of individuals according to their physical and psycho-vegetative traits which until today play a role in complementary medicine. This pilot study aimed at investigating if the concepts of polar constitutional types of anthroposophic medicine and according to Kretschmer can be further clarified using empirical method.; 96 participants, preselected by two polar body mass index (BMI) ranges (17-19.5 kg/m; 2; and 27-31 kg/m; 2; ), were categorized using both classification systems. Anthropometrical measurements were carried out and differences in the autonomic regulation were assessed using a questionnaire. From 12 participants showing a pronounced polar constitutional type, production of reactive oxygen species, proliferation, autophagy, and glucose uptake by lymphocytes, monocytes and granulocytes were measured in vitro.; Correlations between the BMI and the strength of constitutional classification were found for both classification systems. Additionally, a strong correlation between the two systems themselves could be seen. Analysis of the overall questionnaire score of autonomic regulation did not yield significant correlations. However, using a modified 11 item score, reliability (Cronbach α = 0.656) and a differentiation of polar constitutional types was demonstrated (p < 0.001). Regarding the immune function slightly varying levels of reactive oxygen species, autophagy in granulocytes and differences in the strength of inhibition of lymphocyte proliferation by dexamethasone and cyclosporine A were detected. However, most of these in vitro results did not reach significance.; This study represents a first empirical approach toward the classification of anthroposophic constitutional types

Effects of Birch Polypore Mushroom, Piptoporus betulinus (Agaricomycetes), the "Iceman's Fungus", on Human Immune Cells

Piptoporus betulinus, the mushroom that has been carried by Ötzi the "Iceman", has a long tradition of use in medicinal practice for its antiseptic, anticancer, and immune-enhancing properties. With this study, we aimed to investigate the immunomodulatory effects of P. betulinus on primary human immunocompetent cells. The influence of P. betulinus water and methanol extract on apoptosis and necrosis induction of T cells and monocytes was analyzed using annexin V/propidium iodide staining and proliferation of T cells by carboxyfluorescein diacetate succinimidyl ester staining using flow cytometry. The effects on T-cell activation (CD69/CD25) and dendritic cell maturation (CD83, CD86, and CD14) were assessed using flow cytometric analysis of distinct marker expression. Alterations of the secretion of effector mediators interferon (IFN)-γ by T cells and interleukin (IL)-8 by monocytes and dendritic cells were detected by enzyme-linked immunosorbent assay. None of the P. betulinus extracts had a significant influence on apoptosis and necrosis induction, T-cell proliferation, or T-cell activation status, but P. betulinus water extract caused a strong increase in IFN-γ release. The same extract was slightly protective against apoptosis of monocytes and further triggered IL-8 secretion by monocytes and dendritic cells. Moreover, P. betulinus water extract induced dendritic cell maturation. Our results demonstrate the immune-enhancing properties of P. betulinus

Impact of Green Tea Catechin ECG and Its Synthesized Fluorinated Analogue on Prostate Cancer Cells and Stimulated Immunocompetent Cells

Among the known or suspected risk factors, inflammation plays an important role in infectious and non-infectious pathways leading to cancer. Green tea polyphenols have been associated with reducing inflammation and protection against carcinogenesis, especially in prostate cancer. While most of the research in this field, so far, has focussed on epigallocatechin-3-; O; -gallate only, we studied epicatechin-3-; O; -gallate, the second most abundant green tea polyphenol with essential therapeutic potential, to obtain a more detailed understanding of its anti-tumor and anti-inflammatory action. Furthermore, to improve the bioactivity of (-)-epicatechin-3-; O; -gallate, we synthesized a difluoro analogue, called (-)-5,7-difluoro-epicatechin-3-; O; -gallate. Both compounds reduced cell proliferation of human primary inflammatory lymphocytes in an apoptosis-specific fashion, while (-)-5,7-difluoro-epicatechin-3-; O; -gallate had a significantly higher activity compared to the natural product (-)-epicatechin-3-; O; -gallate. Treatment of low-metastatic LNCaP and high-metastatic PC-3 prostate cancer cells with (-)-epicatechin-3-; O; -gallate and (-)-5,7-difluoro-epicatechin-3-; O; -gallate demonstrated a dose-dependent inhibition of cell viability in the low micromolar range. These effects suggest that (-)-epicatechin-3-; O; -gallate and the more effective (-)-5,7-difluoro-epicatechin-3-; O; -gallate could be therapeutically used to inhibit tumorigenesis during initiation, promotion, and progression by diminishing the amount of inflammation due to a reduction of inflammatory lymphocytes. Further studies are needed to prove this in; in vivo; experiments

Ipecac root extracts and isolated circular peptides differentially suppress inflammatory immune response characterised by proliferation, activation and degranulation capacity of human lymphocytes in vitro

Circular peptides are attractive lead compounds for drug development; this study investigates the immunomodulatory effects of defined root powder extracts and isolated peptides (called cyclotides) from Carapichea ipecacuanha (Brot.) L. Andersson ('ipecac'). Changes in the viability, proliferation and function of activated human primary T cells were analysed using flow cytometry-based assays. Three distinct peptide-enriched extracts of pulverised ipecac root material were prepared via C; 18; solid-phase extraction and analysed by reversed-phase HPLC and mass spectrometry. These extracts induced caspase 3/7 dependent apoptosis, thus leading to a suppressed proliferation of activated T cells and a reduction of the number of cells in the G2 phase. Furthermore, the stimulated T cells had a lower activation potential and a reduced degranulation capacity after treatment with ipecac extracts. Six different cyclotides were isolated from C. ipecacuanha and an T cell proliferation inhibiting effect was determined. Furthermore, the degranulation capacity of the T cells was diminished specifically by some cyclotides. In contrast to kalata B1 and its analog T20K, secretion of IL-2 and IFN- γ was not affected by any of the caripe cyclotides. The findings add to our increased understanding of the immunomodulating effects of cyclotides, and may provide a basis for the use of ipecac extracts for immunomodulation in conditions associated with an exessive immune responses

Hapalindoles from the Cyanobacterium Hapalosiphon sp. Inhibit T Cell Proliferation

Novel immunomodulating agents are currently sought after for the treatment of autoimmune diseases and cancers. In this context, a screening campaign of a collection of 575 cyanobacteria extracts for immunomodulatory effects has been conducted. The screening resulted in several active extracts. Here we report the results of subsequent studies on an extract from the cyanobacterium; Hapalosiphon; sp. CBT1235. We identified 5 hapalindoles as the compounds responsible for the observed immunomodulatory effect. These indole alkaloids are produced by several strains of the cyanobacterial family Hapalosiphonaceae. They are known for their anti-infective, cytotoxic, and other bioactivities. Modulation of the activity of human immune cells has not yet been described. The immunomodulatory activity of the hapalindoles was characterized; in vitro; using flow cytometry-based measurements of T cell proliferation after carboxyfluorescein diacetate succinimidyl ester staining, and apoptosis and necrosis induction after annexin V/propidium iodide staining. The most potent compound, hapalindole A, reduced T cell proliferation with an IC; 50; of 1.56 µM, while relevant levels of apoptosis were measurable only at 10-fold higher concentrations. Hapalindole A-formamide and hapalindole J-formamide, isolated for the first time from a natural source, had much lower activity than the nonformylated derivatives while, at the same time, being less selective for antiproliferative over apoptotic effects

Influence of traditionally used Nepalese plants on wound healing and immunological properties using primary human cells in vitro

Ethnopharmacological relevance: The improvement of wound healing has always been an important issue for both ethnopharmacological and modern medical research. In this study, we used state-of-the-art methods to investigate extracts of plants used traditionally in Nepal for more than 1000 years to treat inflammatory injuries. Aim of the study: We focused on the potential of the plant extracts to ameliorate wound healing and to influence immune modulatory properties. Materials and methods: Nine Nepalese plant extracts in three different solvents (methanol, ethyl acetate, petroleum ether) were immunologically characterised. Water-soluble tetrazolium (WST-1) assays and scratch assays were performed to determine their impact on viability and wound healing capacity of human keratinocytes and fibroblasts. Effects on proliferation, viability and function of physiologically relevant anti-CD3 and anti-CD28 stimulated primary human T lymphocytes were assessed using carboxyfluorescein succinimidyl ester (CFSE), annexin V/propidium iodide staining assays and flow cytometry-based surface receptor characterisation. The secretion level of interleukin-2 (IL-2) was analysed with the ELISA technique. Dendritic cells were generated out of peripheral blood mononuclear cells (PBMC) by CD14(+) magnetic bead selection. Flow cytometry-based surface receptor characterisation and ELISA-based technique were used to evaluate the DC activation state and the interleukin-8 (IL-8) secretion level. Results: We demonstrate that an ethyl acetate extract of Bassia longifolia and of Gmelina arborea have anti-inflammatory capacities, indicated by reduced proliferation, inhibition of IL-2 secretion and degranulation capacity of activated human T cells, when compared with adequate concentrations of synthetic positive drug controls. Furthermore, Gmelina arborea improved the wound healing of keratinocytes and fibroblasts and has tendency to increase the secretion of IL-8 by human primary dendritic cells. Conclusion: With this preliminary screening, we offer a scientific basis for the immunomodulatory properties of the two Nepalese medicinal plants Bassia longifolia and Gmelina arborea. However, further detailed studies regarding the responsible compounds are necessary